목차

Contenti
List of Figures ⅴ
Chapter Ⅰ
Purple Perilla extracts allay ER stress in lipid-laden macrophages 1
Abstract 2
Introduction 3
Materials and methods 5
Materials 5
Preparation of purple perilla extracts 5
Preparation of human plasma LDL and oxidation 6
Macrophage culture and viability 6
DNA laddering 7
Hoechst 33258 staining 7
Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay 7
Western blot analysis 8
RT-PCR analysis 8
Immunocytochemical analysis 9
Data analysis 9
Results 10
Inhibitory effect of PPE on ER stress-induced apoptosis 10
Attenuation of ATF6 activation and BiP/GRP78 induction by PPE 12
Blockade of XBP1 induction by PPE 14
Suppressive effect of PPE on oxidized LDL induced-ER stress 17
Retrieval of protein folding activity by PPE 20
Discussion 22
References 25
Chapter Ⅱ
Purple perilla extract and α-asarone enhance cholesterol efflux from lipid-laden macrophages 30
Abstract 31
Introduction 32
Materials and methods 35
Materials 35
Preparation and oxidation of human plasma LDL 35
PPE preparation and identification of α-asarone 36
Cell culture and Oil Red O staining 37
Western blot analysis 37
Cholesterol efflux assay 38
RT-PCR analysis 38
Data analysis 39
Results 40
Blockade of SR-B1 induction and foam cell formation by PPE 40
Up-regulation of ABCA1 and ABCG1 expression and
cholesterol efflux by PPE 42
Augmentation of ABCA1 expression induced by LXR 44
Increase in PPARγ expression by PPE 46
Induction of ABCA1 by PPE-α-asarone 48
Down-regulationg of lipid uptake by PPE-α-asarone 50
Discussion 52
References 56
Chapter III
Dietary compound α-asarone ameliorates ER stress-mediated apoptosis in oxysterol-challenged macrophages 60
Abstract 61
Introduction 62
Materials and methods 65
Materials 65
Macrophage culture and cytotoxicity 65
Terminal deoxynucleotidyl transferase dUTP nick end
labeling (TUNEL) assay 66
Immunocytochemical analysis 66
Western blot analysis 67
Isolation of mitochondrial fractions 67
Dual-luciferase reporter assay 68
Real-time PCR and RT-PCR 68
TRAF2 small interfering RNA transfection 69
Data analysis 69
Results 71
Inhibitory effect of α-asarone on oxysterol-induced apoptosis 71
Attenuation of apoptosome formation by α-asarone 73
Inhibition of caspase 12 activation by α-asarone 76
Diminution of IRE1α downstream signaling by α-asarone 78
Inhibitory effect of α-asarone on XBP1 induction 80
Alleviation of IRE1α-JNK1/2 signaling by α-asarone 82
Suppression of PERK-ATF4 pathway by α-asarone 84
Discussion 86
References 92
Chapter Ⅳ
Dietary compound α-Asarone ameliorates 7β-hydroxycholesterol-induced macrophage injury through blocking eIF2α phosphorylation and prompting beclin-1 dependent autophagy 99
Abstract 100
Introduction 101
Materials and methods 104
Materials 104
Cell culture 104
ER isolation 105
Western blot analysis 105
Autophagolysosome staining 106
RT-PCR and real-time PCR analyses 106
ATG5 small interfering RNA (siRNA) transfection 107
Data analysis 107
Results 108
Inhibition of eIF2α phosphorylation and GADD34 expression by α-asarone 108
Autophagolysosome formation by α-asarone 110
Promotion of autophagy initiation by α-asarone 112
Elevation of LC3 lipidation and p62/SQSTM1 induction by α-asarone 115
α-Asarone potentiation of ATG protein induction
by 7β-hydroxycholesterol 118
Discussion 121
References 126
국문초록 133